The opportunity addressed by this proposal is to provide the scientific and medical communities with the ability to perform rapid, inexpensive, and accurate mutation studies of any protein, using a scaffold based on synthetic genes computationally optimized for DNA self assembly and for expression and translation in an in vitro or in vivo system of choice. Current gene synthesis methods are time-consuming and expensive, and are therefore unsuited to rapid and routine large-scale mutation studies of proteins of biomedical interest. However, whether pathogens occur naturally or are engineered by bioterrorists, mutational variation is one of the primary methods by which they escape the immune system. Indeed, deliberately engineering a pathogen mutant to evade the immune response generated by current vaccines has been identified as one strategy by which a bioterrorist could achieve high penetration into the general population. Easy access to large-scale mutational studies would allow those variations that successfully evade the immune system, and hence render current vaccines ineffective, to be identified, studied, and prepared for in advance. Mutational studies are also one of the primary methods by which biomedical researchers carry out protein sequence-structure studies, and RNA splicing variants are a major source of protein diversity in higher eukaryotes. The ready ability to study mutations and splice variants would advance biology and medicine on many fronts, for example, by supporting pharmacogenomics research, or by enabling protein arrays of the major mutants or splice variants of a protein of interest. Because of worldwide bioterrorism threats and emerging diseases, the development of safe, rapid, and inexpensive manufacturing methods for mutational variants of the genes and proteins of Class A, B & C bioterrorism agents for vaccine development, as well as mutant proteins for all types of biomedical research, is a high national priority.